Ami 1000

All animal studies were approved by the IACUC of Van Andel Research Institute. Subcutaneous and orthotopic [14 (link), 16 (link)] tumor initiation were performed as previously described. The orthotopic tumor growth was measured by bioluminescence signal intensity (BLI) using a small animal optical imager AMI 1000 (Spectral Instruments Imaging, LLC). Dosing with V-4084 and/or erlotinib was delivered once daily by oral gavage for 3 weeks. Vehicles used were 0.5 % MC 400 with 0.05 % Tween 80 (for V-4084) and with 0.5 % (w/v) methyl cellulose (for erlotinib). To determine the effectiveness of treatment, the average tumor size of each group from the last measurement was analyzed with Student’s t test (p < 0.05).

Female six week old athymic-nu/nu mice were purchased from Envigo and handled in accordance with the University of Iowa Animal Care and Use Committee (IACUC). MIA PaCa-2 cells (4 × 10⁵) expressing luciferase and GFP were injected orthotopically directly into the pancreas of mice using ultrasound guidance as previously described [24 (link)]. Cells were suspended in a 1:1 mixture of PBS and Matrigel (Corning, St. Louis, MO) immediately prior to injections to prevent cells from dispersing throughout the abdomen. Tumor formation and growth were monitored weekly through bioluminescent imaging. Mice were given IP injections (200 µl) of a 15 mg/ml solution of VivoGlo^TM luciferin (Promega, Madison, WI). Following injection, mice were anesthetized by isoflurane inhalation and imaged using the AMI-1000 (Spectral Instruments Imaging). Total photon flux (photons per second) was quantified (AMIView software) by placing and area of interest around each mouse. Tumor progression was allowed to continue for fifty days post cancer cell injection.